Despite advances in hygiene, sanitation and the development of vaccines and antibiotics, infectious diseases continue to affect hundreds of millions of people each year with serious health outcomes. Infectious diseases can be transmitted either by air (airborne) or via surfaces (fomites). The toll of infectious disease is further complicated through the evolution of antibiotic-resistant bacteria, while the constant antigenic shift of influenza viruses creates difficulties for vaccine development. Control of these infections remains a challenge and currently relies on interventions that have significant shortcomings, including their own health risks. New, innovative, effective, low cost and most importantly chemical-free, 'green' technologies, possessing fewer drawbacks than the existing ones, are urgently in need in the battle against infections. The investigators have been working on such a novel nanotechnology-based method. It relies on the synthesis of Engineered Water Nanostructures (EWNS) by electrospraying high purity water. Preliminary data indicate that EWNS possess unique physicochemical and biological properties. Most importantly, they are highly mobile and can inactivate bacteria on both surfaces and in the air through damage to their membrane. Here, we plan to assess and optimize EWNS as an alternative, chemical-free method to inactivate viruses in air and on environmental surfaces. The pathogen-EWNS interactions will be investigated using a variety of validated, state-of-the-art analytical methods and biological assays. The specific aims of this project are: AIM1: Development and characterization of a high-throughput EWNS generation platform to study the nano-virus interaction in air and surfaces using relevant bioassay models. The system will be used for the controlled synthesis and property characterization of EWNS. The EWNS generation platform will enable for EWNS property modification (size, surface charge, ROS content, lifetime) and study their effect on the viral inactivation process. AIM 2: Inactivation of aerosolized or surface deposited influenza virus (2009 H1N1) following exposure to EWNS will be assessed and optimized using in vitro and in vivo infectivity assays. The role of EWNS properties and electrospray operational parameters on the inactivation potential and mechanisms will be investigated using state of the art analytical methods. The information generated in these studies will lead to the development of applications of this novel, chemical-free approach for the control of virally transmitted infectious diseases such as Influenza. The proposed project spans disciplines in which our investigators have expertise: Nanoparticle synthesis, characterization and environmental nanotechnology (Demokritou), cellular biology, respiratory pathophysiology, aerobiology and infectious diseases (Kobzik, McDevitt). Such a novel chemical free approach, if successful, will reduce risk of infection and have a beneficial economic and public health impact.